农杆菌介导转化巨桉Eg5影响因素的研究

Study on the factors influencing Agrobacterium tumefaciens mediated transformation of Eucalyptus grandis clone Eg5

以巨桉无性系Eg5叶片为外植体材料,以GUS瞬时表达率和瞬时表达率指数作为评价指标,主要探讨了预培养时间、菌液浓度、侵染时间、共培养基pH、共培养时间和乙酰丁香酮（Acetosyringone,AS）对遗传转化的影响,结果表明预培养3 d、菌液浓度OD600 nm为0.5、接种30~60 min、共培养基pH值为5.8~6.0、共培养基中添加150 mg.L-1AS、菌液中不添加AS、共培养1 d遗传转化效果最佳,初步建立了巨桉Eg5的遗传转化体系。

By taking Eucalyptus grandis clone Eg5 leaves as explants, the GUS gene transient expression efficiency and transient expression efficiency index as evaluation indicators, effects of the pre-culture time, bacterial concentration, infection time, co-culture pH, co-culture time, Acetosyringone （AS） concentration in the bacterium liquid and Acetosyringone concentration in the medium on genetic transformation were investigated. The results show that pre-ctdture 3 days, 0.5 of OD600nm, 30-60 rain of infection time, 5.8,-6.0 of co-culture pH, 150 mg·L-t AS of the medium, and no AS in the bacterium liquid, lday of co-culture time, the best transformation effects were achieved. Thus, an effective system for Agrobacterium-mediated transformation of Eucalyptus grandis clone Eg5 was established.